External traditional Chinese medicine preparation for internal arteriovenous fistula stenosis and preparation method thereof

Abstract

The invention relates to the field of traditional Chinese medicine preparations, in particular to an external traditional Chinese medicine preparation for internal arteriovenous fistula stenosis, which is prepared from the following components in parts by weight: 18 to 22 parts of radix salviae miltiorrhizae, 4 to 6 parts of radix angelicae sinensis, 5 to 10 parts of lumbricus, 8 to 12 parts of frankincense, 8 to 12 parts of myrrh, 20 to 30 parts of radix astragali seu hedysari and 15 to 20 parts of caulis spatholobi. Internal fistula stenosis caused by internal fistula vascular calcium deposition can be effectively treated and prevented.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of traditional Chinese medicine preparations, in particular to an external traditional Chinese medicine preparation for internal arteriovenous fistula stenosis and a preparation method thereof. BACKGROUND

[0002] Patients with end-stage renal disease need to have a fistula for hemodialysis, and an internal arteriovenous fistula is generally preferred due to low risk of infection, long use time, and few complications. However, the internal arteriovenous fistula may be stenotic due to factors such as the patient's own vascular condition, damage caused by surgery, and improper postoperative care, which affects the dialysis adequacy of the patient.

[0003] Some traditional Chinese medicine preparations are coated on the fistula of the patient, which can improve the vascular condition of the patient and reduce the occurrence of internal fistula stenosis, for example, a traditional Chinese medicine composition for treating postoperative complications of internal arteriovenous fistula disclosed in CN105232673A, which is prepared by compounding papaya, rhodiola, white peony root, sedum lineare, trumpet flower, Chinese albizia, melia toosendan, astragalus, wintergreen, salvia miltiorrhiza, caesalpinia and randia.

[0004] However, the above traditional Chinese medicine composition has relatively weak treatment effect on one of the factors of internal fistula vascular stenosis, i.e., vascular calcium deposition. SUMMARY

[0005] In order to more effectively treat vascular calcium deposition of the patient, the present application provides an external traditional Chinese medicine preparation for internal arteriovenous fistula stenosis and a preparation method thereof.

[0006] In a first aspect, the external traditional Chinese medicine preparation for internal arteriovenous fistula stenosis provided by the present application adopts the following technical solution.

[0007] An external traditional Chinese medicine preparation for internal arteriovenous fistula stenosis includes the following components by weight: 18-22 parts of salvia miltiorrhiza, 4-6 parts of angelica sinensis, 5-10 parts of earthworm, 8-12 parts of frankincense, 8-12 parts of myrrh, 20-30 parts of astragalus, and 15-20 parts of caesalpinia.

[0008] By adopting the above technical solution, salvia miltiorrhiza and angelica sinensis cooperate to activate blood circulation to remove blood stasis, earthworm, frankincense and myrrh have anti-inflammatory and analgesic effects, astragalus and caesalpinia promote vascular repair, and the beta-sitosterol in angelica sinensis significantly improves the calcium ion regulation efficiency of salvianolic acid B in salvia miltiorrhiza by strengthening the function of TRPV1 channel, and in combination with the precise decomposition of fibrin network by earthworm enzyme in earthworm, the matrix environment of pathological calcium deposition is eliminated, forming a triple anti-calcification mechanism, which effectively prevents and treats internal fistula vascular calcium deposition and reduces the possibility of internal fistula vascular stenosis.

[0009] Optionally, 5-8 parts of rhizoma dipsaci are further included by weight.

[0010] By adopting the technical scheme, dioscin in Rhizoma Dioscoreae can activate TRPV4 channel to increase calcium ion efflux capacity, and effectively reduce the possibility of vascular calcium deposition of internal fistula.

[0011] Optionally, 3-5 parts of Uncaria by weight are further included.

[0012] By adopting the technical scheme, isorhynchophylline in Uncaria can inhibit ROCK-MYPT1 phosphorylation, regulate vascular tension, and relieve vascular spasm.

[0013] Optionally, the following components are included by weight: 20 parts of Salvia miltiorrhiza, 5 parts of Angelica sinensis, 7 parts of Pheretima, 10 parts of Olibanum, 10 parts of Myrrha, 25 parts of Astragalus membranaceus, 18 parts of Spatholobus suberectus, 6 parts of Rhizoma Dioscoreae, and 4 parts of Uncaria.

[0014] By adopting the technical scheme, the preferred proportion of each component can achieve a better treatment and prevention effect on vascular calcium deposition.

[0015] In a second aspect, the application provides a preparation method of an external use traditional Chinese medicine preparation for internal arteriovenous fistula stenosis.

[0016] A preparation method of an external use traditional Chinese medicine preparation for internal arteriovenous fistula stenosis, which is used for preparing the external use traditional Chinese medicine preparation for internal arteriovenous fistula stenosis, and specifically includes the following steps.

[0017] Step 1, pretreating or processing each component medicine;

[0018] Step 2, extracting effective components from each processed medicine;

[0019] Step 3, adding the Salvia miltiorrhiza extract, Angelica sinensis extract, and Uncaria extract, as well as soybean phospholipid and cholesterol, into an organic solvent for mixing, then removing the organic solvent, adding the Astragalus membranaceus extract and Spatholobus suberectus extract buffer for hydration, and then filtering to obtain a nanoparticle suspension;

[0020] Step 4, preparing a gel system, adding the nanoparticle suspension first, then adding the Pheretima extract, Olibanum extract, Myrrha extract, and Rhizoma Dioscoreae extract and stirring.

[0021] By adopting the technical scheme, the nanocarrier system and the gel matrix are integrated to improve the transdermal absorption rate.

[0022] Optionally, in the step two, the Salvia miltiorrhiza and Angelica sinensis are mixed in a certain proportion, then citric acid buffer is added and ultrasonic extraction is performed, then the filtrate is separated and purified through a resin column, then the eluate is collected and concentrated under reduced pressure to obtain a mixed extract of Salvia miltiorrhiza and Angelica sinensis.

[0023] By adopting the technical scheme, the effective components in the Danshen and the Angelica are efficiently enriched, and the β-sitosterol in the Angelica and the salvianolic acid B in the Danshen are dissolved in the citric acid buffer solution for extraction and purification, so that the oxidation of the β-sitosterol and the salvianolic acid B can be effectively prevented.

[0024] Optionally, in the second step, the earthworms are added into the physiological saline, homogenized, and then subjected to ultrasonic crushing, and then centrifuged to obtain supernatant, and then subjected to column chromatography purification, and then the components with corresponding molecular weight are collected and freeze-dried to obtain the earthworm active peptide powder.

[0025] By adopting the technical scheme, the high-activity peptide segment molecules with corresponding molecular weight can be extracted from the earthworm polypeptide molecules with wide molecular weight, so that the prepared traditional Chinese medicine preparation can more effectively improve the activity of blood vessel neogenesis.

[0026] Optionally, in the second step, the Rhizoma Dioscoreae Nipponicae is added into the phosphate buffer solution, and then β-glucosidase is added and subjected to oscillation reaction, and then the enzyme is inactivated by heating, and then the supernatant is obtained by centrifugation and filtration, and then the anhydrous ethanol is added after the reduced pressure concentration, and then the Rhizoma Dioscoreae Nipponicae enzymatic powder is obtained by resin purification and spray drying.

[0027] By adopting the technical scheme, the original saponin in the Rhizoma Dioscoreae Nipponicae with low bioavailability is directionally converted into dioscin, so as to effectively improve the bioavailability.

[0028] Optionally, in the second step, the Uncaria is added into the hydrochloric acid solution and subjected to ultrasonic extraction, and then the extraction liquid is extracted for several times, and then the organic phase is combined and dehydrated, and then the solvent is recovered and dissolved in the water phase again, and then the pH gradient separation is performed by adopting the countercurrent distribution chromatography, and then the alkaloid main peak in the corresponding interval is collected, and then the Uncaria alkaloid crystal powder is obtained after concentration and drying.

[0029] By adopting the technical scheme, the Uncaria alkaloid with similar chemical structure to the isorhynchophylline in the Uncaria is effectively separated, so as to improve the purity of the isorhynchophylline.

[0030] Optionally, in the second step, the Astragalus and the Spatholobus subopposite leaf are mixed in proportion, and then the purified water and the cellulase are added for enzymolysis, and then the enzyme is inactivated by boiling and centrifugation after the enzymolysis is completed, and then the supernatant is filtered and freeze-dried to obtain the Astragalus and Spatholobus subopposite leaf composite extract.

[0031] By adopting the technical scheme, the problem of poor compatibility and high precipitation rate of the Astragalus polysaccharide and the Spatholobus subopposite leaf flavone is solved, and the solubility of the composite is effectively improved, and the extraction efficiency and the purity are improved.

[0032] In summary, the present application includes at least the following beneficial effects:

[0033] 1、Angelica sinensis β-sitosterol can significantly improve the calcium ion regulation efficiency of Danshensu in Salvia miltiorrhiza by strengthening the function of TRPV1 channel, and combined with the precise decomposition of fibrin network by earthworm enzyme in Pheretima, the matrix environment of pathological calcium deposition is eliminated, forming a triple anti-calcification mechanism, effectively preventing and treating internal fistula vascular calcification, and reducing the possibility of internal fistula vascular stenosis;

[0034] 2、Dioscorea saposhnikovii in Rhizoma Dioscoreae can activate TRPV4 channel to increase calcium ion efflux capacity, effectively inhibit internal fistula vascular calcification;

[0035] 3、Uncaria rhynchophylline in Uncaria can inhibit ROCK-MYPT1 phosphorylation, regulate vascular tension, relieve vascular spasm, and reduce oxidative stress, thereby inhibiting vascular calcification;

[0036] 4、Nanometer drug delivery system and gel matrix are integrated to improve transdermal absorption rate;

[0037] 5、Angelica sinensis β-sitosterol and Danshensu in Salvia miltiorrhiza are dissolved in citric acid buffer for extraction and purification, which can effectively prevent oxidation of β-sitosterol and Danshensu;

[0038] 6、Extract high-activity peptide molecules with corresponding molecular weight from Pheretima polypeptide molecules with wide molecular weight, so that the prepared traditional Chinese medicine preparation can more effectively improve the activity of vascularization;

[0039] 7、Biological utilization of low Dioscorea saposhnikovii in Rhizoma Dioscoreae is enzymatically converted into dioscin, which effectively improves the bioavailability;

[0040] 8、Uncaria rhynchophylline with similar chemical structure to Uncaria rhynchophylline is effectively separated to improve the purity of Uncaria rhynchophylline;

[0041] 9、Solve the problem of poor compatibility and high precipitation rate of Astragalus polysaccharide and Morinda officinalis flavonoids, effectively improve the solubility of the compound, and improve the extraction efficiency and purity. DETAILED DESCRIPTION

[0042] The application discloses an external use traditional Chinese medicine preparation for arteriovenous fistula stenosis, which comprises the following components in parts by weight: 18-22 parts of Salvia miltiorrhiza, 4-6 parts of Angelica sinensis, 5-10 parts of Pheretima, 8-12 parts of Olibanum, 8-12 parts of Myrrha, 20-30 parts of Astragalus membranaceus, 15-20 parts of Morinda officinalis, 5-8 parts of Rhizoma Dioscoreae, and 3-5 parts of Uncaria.

[0043] The application also discloses a preparation method of the external use traditional Chinese medicine preparation for arteriovenous fistula stenosis.

[0044] Step 1, pretreat or process each component of medicinal materials, specifically as follows.

[0045] Salvia miltiorrhiza: Take dried Salvia miltiorrhiza medicinal materials, remove impurities and residual stems and leaves, soak in clean water until the heart is transparent, cut into 3-5 mm thick slices, place in a hot air circulation oven and dry at 60°C until the moisture is less than or equal to 8%, then pulverize with a universal pulverizer and pass through a 40 mesh sieve, and finally seal and store for future use.

[0046] Angelica sinensis: Select whole Angelica sinensis without insect damage, spray with yellow wine and mix well, soak for 2 hours, then place in a frying machine and fry at 140°C for 15 minutes until the surface is dark yellow, then spread and cool, pulverize and pass through a 40 mesh sieve, and finally sterilize by irradiation for future use.

[0047] Radix astragali: Take Radix astragali decoction pieces, add 10 times the amount of purified water and decoct twice, the first time for 1.5 hours and the second time for 1 hour, filter both times, combine the two filtrates, reduce the filtrate to a thick paste under vacuum, then dry the thick paste under vacuum at 65°C and 0.08 MPa with water as the reference material, and finally pulverize and pass through an 80 mesh sieve to obtain Radix astragali extract.

[0048] Spatholobus suberectus: Put Spatholobus suberectus segments into a multifunctional extraction tank, add 8 times the amount of 70% ethanol, and reflux extract 3 times for 1 hour each time, combine the three extraction liquids, then purify the extraction liquid with macroporous resin D101, use 50% ethanol as the eluent, collect the eluate and concentrate to remove the alcohol taste, and finally spray dry to obtain a brownish red powder with a moisture content of less than or equal to 5%.

[0049] Earthworm: Take earthworm decoction pieces, use sand scalding method, i.e. stir-fry river sand at 240°C until the surface is dark brown and the cross section is brownish yellow, then pulverize and pass through a 60 mesh sieve, sterilize by irradiation and store for future use.

[0050] Frankincense and myrrh: Take frankincense and myrrh, mix with rice vinegar, and stir-fry over low heat until the surface is shiny and the irritating gas is gone, then pulverize and pass through a 50 mesh sieve and store in a sealed container.

[0051] Dipsacus asper: Remove the outer skin of fresh Dipsacus asper, cut into segments about 5 cm long, perform microwave rapid enzyme inactivation treatment for 90s at 800W power, then dry at 60°C, and finally pulverize and pass through a 40 mesh sieve.

[0052] Uncaria rhynchophylla: Take Uncaria rhynchophylla stems with hooks, perform supercritical CO2 degreasing at a pressure of 25 MPa and a temperature of 40°C, and finally pulverize and pass through a 60 mesh sieve and store at low temperature.

[0053] Step 2: Extract the effective components from each treated medicinal material, as follows.

[0054] Danshen and Angelica: Danshen coarse powder and processed Angelica powder are mixed in proportion, 10 times of citric acid buffer solution with pH 3.0 is added, and ultrasonic extraction is carried out at 50°C with a frequency of 40 kHz and a power of 800 W. The ultrasonic extraction is carried out for 3 times and each time for 30 min. The combined extract is filtered by plate frame, and the filtrate is passed through an HPD-300 macroporous resin column. First, 3 times of column volume of pure water is used to elute impurities, and then 70% concentration ethanol is used to elute target components. The eluate is concentrated under reduced pressure to obtain a mixed extract of Danshen and Angelica.

[0055] Huangqi and Caobuqie: Huangqi thick extract and Caobuqie alcohol extract are mixed in proportion, 5 times of purified water is added for re-dissolution, cellulase is added, and the proportion of cellulase in the mixture is 0.1% after addition. Then, enzymatic hydrolysis is carried out at 45°C for 2h. After boiling to inactivate the enzyme, centrifugation is carried out. The supernatant is subjected to ultrafiltration to remove macromolecular impurities. Finally, freeze-drying is carried out to obtain a yellow-brown Caobuqie and Huangqi composite extract.

[0056] Dilong: Sand scalded Dilong powder is added with 8 times of normal saline, and high pressure homogenization is carried out at 120 MPa. Then, ultrasonic crushing is carried out at a frequency of 20 kHz for 30 min. The supernatant is obtained by centrifugation, and is subjected to Sephadex G-25 column chromatography purification. The components with a molecular weight of 3-10 kDa are collected, and freeze-drying is carried out to obtain Dilong active peptide powder.

[0057] Muxiang and Muyang: Vinegar Muxiang and Muyang powder are mixed in proportion, 5 times of 95% concentration ethanol is added and cold soaked for 24h. Reflux extraction is carried out for 2 times and each time for 1 hour. The two times of extract are combined and the ethanol is recovered under reduced pressure. The residue is subjected to silica gel column chromatography. The fractions containing mastic acid and bisabolane are collected. Finally, rotary evaporation is carried out to concentrate to a paste to obtain Muxiang and Muyang resin extract.

[0058] Chuanshanyang: Chuanshanyang powder is added with 5 times of phosphate buffer solution with pH 5.0, and β-glucosidase is added. Each gram of raw material contains 20U of β-glucosidase. The constant temperature oscillation reaction is carried out at 50°C for 2h. Then, the temperature is quickly raised to 90°C and maintained for 10 min to inactivate the enzyme. The supernatant is obtained by centrifugation and is filtered by a ceramic membrane with a pore size of 0.45μm. The filtrate is concentrated under reduced pressure to 1 / 3 of the original volume. Then, 3 times of anhydrous ethanol is added to precipitate polysaccharide impurities and is filtered again. The filtrate is subjected to macroporous resin purification and is spray dried to obtain Chuanshanyang enzymatic powder.

[0059] Uncaria: defatted Uncaria powder is ultrasonically extracted with 8 times the amount of 0.5% hydrochloric acid solution, the extract is adjusted to pH 10.0, and then extracted with chloroform for 3 times, the organic phases of the three extractions are combined and dehydrated with anhydrous sodium sulfate, then rotary evaporation is used to recover the solvent and obtain crude alkaloids, then the crude alkaloids are dissolved in a water phase with pH 3.0, and then pH gradient separation is performed by countercurrent distribution chromatography, the main peak of alkaloids in the pH 7.0-8.0 interval is collected, and then concentrated and dried to obtain white Uncaria alkaloid crystal powder.

[0060] Step 3, the extract of Salvia miltiorrhiza and Angelica sinensis, Uncaria alkaloid crystal powder, soybean phospholipid and cholesterol are added to the organic solvent in proportion and mixed, the organic solvent can be a chloroform-methanol solution configured in a ratio of 9:1, then rotary evaporation is used to remove the organic solvent to form a uniform lipid film, then the Astragalus and Millettia pinnata complex extract and the phosphate buffer solution with pH 7.4 are added to the lipid film for hydration reaction for 30 min, then ultrasonic treatment is performed at a power of 100 w for 5 min, then filtered through a 0.22 μm microporous filter to obtain a drug-loaded nanoparticle suspension.

[0061] Step 4, a gel system is prepared, the nanoparticle suspension, earthworm active peptide powder, frankincense and myrrh resin extract, and Dipsacus asper enzymatic powder are added to the gel system in proportion, then constant temperature stirring is maintained at 40°C for 40 min, then a high-pressure homogenizer is used to run at a speed of 10000 rpm for 10 min to make the system uniformly dispersed, then vacuum degassing is performed, then filled into an aluminum tube, and then irradiation sterilization is performed to obtain an orange-yellow translucent gel preparation.

[0062] The preparation of the gel system is specifically as follows: the deacetylated gelling gum is dispersed in purified water, dissolved by stirring at 80°C for 1 h, then chitosan with a deacetylation degree greater than or equal to 95% is added, and stirring is continued until completely dissolved, then the temperature is lowered to 45°C for standby.

[0063] The following is described in detail through multiple examples and comparative examples.

[0064] Example 1:

[0065] An external use traditional Chinese medicine preparation for treating arteriovenous fistula stenosis, which comprises the following components in parts by weight: Salvia miltiorrhiza 18 parts, Angelica sinensis 4 parts, earthworm 5 parts, frankincense 8 parts, myrrh 8 parts, Astragalus 20 parts, Millettia pinnata 15 parts, Dipsacus asper 5 parts, and Uncaria 3 parts.

[0066] A preparation method of an external use traditional Chinese medicine preparation for treating arteriovenous fistula stenosis, which specifically comprises the following steps.

[0067] Step 1, pretreatment or processing of each component of medicinal material.

[0068] Step 2, extraction of effective components from each treated medicinal material.

[0069] Danshen and Danggui: Danshen coarse powder and processed Danggui powder are mixed in proportion, 10 times the amount of citric acid buffer solution with pH 3.0 is added, and ultrasonic extraction is carried out at 50°C with a frequency of 40 kHz and a power of 800 W. The ultrasonic extraction is carried out for 3 times and each time for 30 min. The combined extract is filtered by plate and frame, and the filtrate is passed through an HPD-300 macroporous resin column. First, 3 times the column volume of pure water is used to elute impurities, and then 70% concentration ethanol is used to elute target components. The eluate is concentrated under reduced pressure to obtain a mixed extract of Danshen and Danggui.

[0070] Huangqi and Jixueteng: Huangqi thick extract and Jixueteng alcohol extract are mixed in proportion, 5 times the amount of purified water is added for re-dissolution, cellulase is added, and the proportion of cellulase in the mixture is 0.1% after addition. Then, enzymatic hydrolysis is carried out at 45°C for 2h. After boiling to inactivate the enzyme, centrifugation is performed. The supernatant is subjected to ultrafiltration to remove macromolecular impurities. Finally, freeze-drying is performed to obtain a yellow-brown Huangqi and Jixueteng composite extract.

[0071] Dilong: Sand scalded Dilong powder is added with 8 times the amount of normal saline, and high-pressure homogenization is carried out at 120 MPa. Then, ultrasonic crushing is carried out at a frequency of 20 kHz for 30 min. The supernatant is obtained by centrifugation, and is subjected to Sephadex G-25 column chromatography purification. The components with a molecular weight of 3-10 kDa are collected, and freeze-drying is performed to obtain Dilong active peptide powder.

[0072] Muxiang and Muyan: Vinegar Muxiang and Muyan powder are mixed in proportion, 5 times the amount of 95% concentration ethanol is added and cold soaked for 24h. Reflux extraction is carried out for 2 times and each time for 1 hour. The two times of extract are combined and the ethanol is recovered under reduced pressure. The residue is subjected to silica gel column chromatography. The fractions containing mastic acid and muurolane are collected. Finally, rotary evaporation is carried out to concentrate to a paste to obtain Muxiang and Muyan resin extract.

[0073] Chuanshanyulong: Chuanshanyulong powder is added with 5 times the amount of phosphate buffer solution with pH 5.0. β-glucosidase is added, and each gram of raw material contains 20U of β-glucosidase. The reaction is carried out at 50°C for 2h. Then, the temperature is quickly raised to 90°C and maintained for 10 min to inactivate the enzyme. The supernatant is obtained by centrifugation and is filtered by a ceramic membrane with a pore size of 0.45μm. The filtrate is concentrated under reduced pressure to 1 / 3 of the original volume. Then, 3 times the amount of anhydrous ethanol is added to precipitate polysaccharide impurities and filter again. The filtrate is subjected to macroporous resin purification and then spray-dried to obtain Chuanshanyulong enzymatic powder.

[0074] Uncaria: defatted Uncaria powder is ultrasonically extracted with 8 times the amount of 0.5% hydrochloric acid solution, the extract is adjusted to pH 10.0, and then extracted with chloroform for 3 times, the organic phases of the three extractions are combined and dehydrated with anhydrous sodium sulfate, then rotary evaporation is used to recover the solvent and obtain crude alkaloids, then the crude alkaloids are dissolved in a water phase with pH 3.0, and then pH gradient separation is performed by countercurrent distribution chromatography, the main peak of alkaloids in the pH 7.0-8.0 interval is collected, and then concentrated and dried to obtain white Uncaria alkaloid crystal powder.

[0075] Step 3, the extract of Salvia miltiorrhiza and Angelica sinensis, Uncaria alkaloid crystal powder, soybean phospholipid and cholesterol are mixed in proportion in an organic solvent, which can be a chloroform-methanol solution configured in a ratio of 9:1, then the organic solvent is removed by rotary evaporation to form a uniform lipid film, then the Astragalus and Millettia pachycarpa complex extract and a mixture of phosphate buffer with pH 7.4 are added to the lipid film for a hydration reaction of 30 min, then ultrasonic treatment is performed at a power of 100 w for 5 min, then filtered through a 0.22 μm microporous filter to obtain a drug-loaded nanoparticle suspension.

[0076] Step 4, a gel system is prepared, and the nanoparticle suspension, earthworm active peptide powder, frankincense and myrrh resin extract, and Dipsacus asper enzymatic powder are added in proportion to the gel system, then constant temperature stirring at 40℃ is maintained for 40 min, then a high-pressure homogenizer is used to run at a speed of 10000 rpm for 10 min to make the system uniformly dispersed, then vacuum degassing is performed, then filled into an aluminum tube, and then irradiation sterilization is performed to obtain an orange-yellow translucent gel preparation.

[0077] The preparation of the gel system is specifically as follows: the deacetylated gellan gum is dispersed in purified water, dissolved by stirring at 80℃ for 1 h, then chitosan with a deacetylation degree greater than or equal to 95% is added, and stirring is continued until complete dissolution, then the temperature is lowered to 45℃ for standby.

[0078] Example Two:

[0079] The difference between Example One and Example Two is that the external use traditional Chinese medicine preparation for arteriovenous fistula stenosis comprises the following components in parts by weight: Salvia miltiorrhiza 20 parts, Angelica sinensis 5 parts, earthworm 7 parts, frankincense 10 parts, myrrh 10 parts, Astragalus 25 parts, Millettia pachycarpa 18 parts, Dipsacus asper 6 parts, and Uncaria 4 parts.

[0080] Example Three:

[0081] The difference between Example One and Example Three is that the external use traditional Chinese medicine preparation for arteriovenous fistula stenosis comprises the following components in parts by weight: Salvia miltiorrhiza 22 parts, Angelica sinensis 6 parts, earthworm 10 parts, frankincense 12 parts, myrrh 12 parts, Astragalus 30 parts, Millettia pachycarpa 20 parts, Dipsacus asper 8 parts, and Uncaria 5 parts.

[0082] Example Four:

[0083] The difference between the example one and the example two is that the external use traditional Chinese medicine preparation for treating arteriovenous fistula stenosis comprises the following components in parts by weight: Danshen 20 parts, Danggui 5 parts, Dilong 7 parts, Muxiang 10 parts, Muyan 10 parts, Huangqi 25 parts, Jixueteng 18 parts.

[0084] Example five:

[0085] The difference between the example one and the example five is that the external use traditional Chinese medicine preparation for treating arteriovenous fistula stenosis comprises the following components in parts by weight: Danshen 20 parts, Danggui 5 parts, Dilong 7 parts, Muxiang 10 parts, Muyan 10 parts, Huangqi 25 parts, Jixueteng 18 parts, Chuanshanlong 6 parts.

[0086] Comparative example one:

[0087] The difference between the example one and the example five is that the external use traditional Chinese medicine preparation for treating arteriovenous fistula stenosis comprises the following components in parts by weight: Danshen 20 parts, Danggui 5 parts, Dilong 7 parts, Muxiang 10 parts, Muyan 10 parts, Huangqi 25 parts, Jixueteng 18 parts, Chuanshanlong 6 parts.

[0088] A preparation method of an external use traditional Chinese medicine preparation for treating arteriovenous fistula stenosis, specifically comprising the following steps.

[0089] Step 1, according to the prescription, the following components are weighed: Zihuadeng, Qianliugong, Baizhen, Baixianpi, Aloe, Xieshangcao, Zixue Guanzhong, Jingjie, Dijincao, are added with water and decoct twice (8 times the amount of water is added in the first time, decoct for 2 hours; 6 times the amount of water is added in the second time, decoct for 1.5 hours), the filtrates are combined, concentrated to the relative density of 1.2 (50℃) of the extract, dried at 80℃ under reduced pressure, pulverized, and passed through a 100 mesh sieve for standby use;

[0090] Step 2, Lingshiaohua, Huhuopi and Shidan are pulverized, passed through a 100 mesh sieve, and then mixed with the traditional Chinese medicine extract powder.

[0091] Comparative example two:

[0092] The difference between the example one and the example five is that in the preparation method of the external use traditional Chinese medicine preparation for treating arteriovenous fistula stenosis, step 3 is that Danshen and Danggui mixed extract, Huangqi and Jixueteng compound extract, Dilong active peptide powder, Muxiang and Muyan resin extract, Chuanshanlong enzymatic powder and Gouteng alkaloid crystal powder are mixed, 2% of azone (penetration agent) is added in total weight, and then 75% concentration of ethanol solution is used for constant volume to 100%, stirred and mixed, and then divided and packaged for use.

[0093] The traditional Chinese medicine preparations obtained in each example and comparative example are subjected to the following experiments.

[0094] The SD rat carotid-jugular internal fistula model was established, and the fistula was smeared daily from the fifth week after the operation for six weeks. On the basis of five example groups and two comparative example groups, a blank control group, a model control group and a positive control group were added, and the sample size of each group was 10. The blank control group was smeared with normal saline to the unmodeled healthy rats. The model control group was smeared with a gel system without drugs to the modeled rats. The positive control group was smeared with a rapamycin gel to the modeled rats. The smearing amount of the positive control group was 1 mg / cm², and the smearing amount of each example and comparative example was 0.5 g / cm².

[0095] After six weeks of continuous smearing, the internal fistula blood vessel tissue was taken, the calcium deposition amount was detected by the o-cresol red complex ketone method, the osteogenic differentiation marker RUNX2 expression was detected by Western Blot, the 24-hour cumulative amount of transdermal penetration was determined by the Franz diffusion cell, and the drug retention amount in the dermis was detected by LC-MS / MS.

[0096] The specific experimental results are shown in the following table.

[0097]

[0098] As can be seen from Examples 1 to 5 and Comparative Example 1, β-sitosterol in angelica can significantly improve the calcium ion regulation efficiency of salvianolic acid B in salvia miltiorrhiza by strengthening the function of TRPV1 channel, and in combination with earthworm enzyme in earthworm that can precisely decompose fibrin network to eliminate the matrix environment of pathological calcium deposition, to form a triple anti-calcification mechanism. In addition, dioscin in rhizoma dioscoreae can activate the TRPV4 channel to increase the calcium ion efflux capacity, effectively inhibit the internal fistula blood vessel calcium deposition, and iso-hooking alkaline in Uncaria can inhibit ROCK-MYPT1 phosphorylation, regulate blood vessel tension, relieve vascular spasm, and reduce oxidative stress, thereby inhibiting vascular calcium deposition.

[0099] As can be seen from Examples 1 to 5 and Comparative Example 2, the use of a nano drug delivery system and a gel matrix integration can effectively improve the transdermal absorption rate, and the proportion of each component in the medicament will also affect the lipophilicity and drug release of the medicament to some extent. Therefore, the proportion in Example 2 is a preferred proportion of the present application.

[0100] The above are preferred embodiments of the present application, which do not limit the protection scope of the present application, therefore: any equivalent changes made on the basis of the structure, shape and principle of the present application should be covered within the protection scope of the present application.

Claims

1. A Chinese medicinal preparation for external use for arteriovenous fistula stenosis, characterized by: The composition comprises the following components by weight: 18-22 parts of salvia miltiorrhiza, 4-6 parts of angelica sinensis, 5-10 parts of earthworm, 8-12 parts of frankincense, 8-12 parts of myrrh, 20-30 parts of astragalus, and 15-20 parts of millettia reticulata.

2. The external-use Chinese medicinal preparation for arteriovenous fistula stenosis according to claim 1, characterized in that: The invention also includes 5-8 parts of Rhizoma Dioscoreae by weight.

3. The external-use Chinese medicinal preparation for arteriovenous fistula stenosis according to claim 2, characterized in that: The invention also includes 3-5 parts of Uncaria rhynchophylla by weight.

4. The external-use Chinese medicinal preparation for arteriovenous fistula stenosis according to claim 3, characterized in that: The composition includes the following components by weight: 20 parts of salvia miltiorrhiza, 5 parts of angelica sinensis, 7 parts of earthworm, 10 parts of frankincense, 10 parts of myrrh, 25 parts of astragalus, 18 parts of millettia reticulata, 6 parts of rhizoma dichotomae, and 4 parts of Uncaria rhynchophylla.

5. A method for preparing a traditional Chinese medicine preparation for external use for treating arteriovenous fistula stenosis, comprising preparing the traditional Chinese medicine preparation for external use for treating arteriovenous fistula stenosis according to any one of claims 1 to 4, characterized in that: The specific steps include: Step 1: pre-treating or processing each component medicinal material; Step 2, extracting effective ingredients from each processed medicinal material; Step 3, adding the extracts of Danshen, Angelica and Uncaria rhynchophylla, soybean lecithin and cholesterol into an organic solvent and mixing them, then removing the organic solvent, adding a buffer solution of Astragalus and Millettia reticulata extracts to hydrate, and then filtering to obtain a nanoparticle suspension; Step 4: prepare a gel system by first adding the nanoparticle suspension, and then adding the extracts of earthworm, frankincense, myrrh and rhizome of radix scholerae and stirring.

6. The method for preparing an external-use Chinese medicinal preparation for arteriovenous fistula stenosis according to claim 5, characterized in that: In the step 2, Danshen and Angelica are mixed in proportion, and then citric acid buffer is added and ultrasonic extraction is performed. The mixture is then filtered, and the filtrate is passed through a resin column for separation and purification. The eluate is then collected and concentrated under reduced pressure to obtain a mixed extract of Danshen and Angelica.

7. The method for preparing an external-use Chinese medicinal preparation for arteriovenous fistula stenosis according to claim 5, characterized in that: In the second step, the earthworm is added to physiological saline, homogenized and then ultrasonically crushed, and then centrifuged to obtain the supernatant, which is then purified by column chromatography, and the corresponding molecular weight components are collected and freeze-dried to obtain the earthworm active peptide powder.

8. The method for preparing an external-use Chinese medicinal preparation for arteriovenous fistula stenosis according to claim 5, characterized in that: In the second step, the Herba Dioscoreae is added to a phosphate buffer solution, β-glucosidase is added and the reaction is shaken, the enzyme is inactivated by heating, the supernatant is collected by centrifugation and filtered, the supernatant is concentrated under reduced pressure, anhydrous ethanol is added, and the Herba Dioscoreae is purified by resin and spray-dried to obtain Herba Dioscoreae enzymatic hydrolysis powder.

9. The method for preparing an external-use Chinese medicinal preparation for arteriovenous fistula stenosis according to claim 5, characterized in that: In the step 2, Uncaria rhynchophylla is added to a hydrochloric acid solution and ultrasonically extracted, the extract is extracted several times, the organic phases are combined and dehydrated, the solvent is recovered and then dissolved in an aqueous phase, and pH gradient separation is performed using countercurrent distribution chromatography, the main peak of the alkaloids in the corresponding interval is collected, and the Uncaria rhynchophylla alkaloid crystalline powder is obtained after concentration and drying.

10. The method for preparing an external-use Chinese medicinal preparation for arteriovenous fistula stenosis according to claim 5, characterized in that: In the step 2, astragalus and Spatholobus suberectus are mixed in proportion, purified water and cellulase are added for enzymolysis, the enzyme is inactivated by boiling after the enzymolysis is completed, and the mixture is centrifuged, the supernatant is filtered and freeze-dried to obtain the astragalus and Spatholobus suberectus composite extract.

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